Cyclodepsipeptide Biosynthesis in Hypocreales Fungi and Sequence Divergence of The Non-Ribosomal Peptide Synthase Genes

Fungi from the Hypocreales order synthesize a range of toxic non-ribosomal cyclic peptides with antimicrobial, insecticidal and cytotoxic activities. Entomopathogenic Beauveria, Isaria and Cordyceps as well as phytopathogenic Fusarium spp. are known producers of beauvericins (BEAs), beauvenniatins (BEAEs) or enniatins (ENNs). The compounds are synthesized by beauvericin/enniatin synthase (BEAS/ESYN1), which shows significant sequence divergence among Hypocreales members. We investigated ENN, BEA and BEAE production among entomopathogenic (Beauveria, Cordyceps, Isaria) and phytopathogenic (Fusarium) fungi; BEA and ENNs were quantified using an LC-MS/MS method. Phylogenetic analysis of partial sequences of putative BEAS/ESYN1 amplicons was also made. Nineteen fungal strains were identified based on sequence analysis of amplified ITS and tef-1α regions. BEA was produced by all investigated fungi, with F. proliferatum and F. concentricum being the most efficient producers. ENNs were synthesized mostly by F. acuminatum, F. avenaceum and C. confragosa. The phylogeny reconstruction suggests that ancestral BEA biosynthesis independently diverged into biosynthesis of other compounds. The divergent positioning of three Fusarium isolates raises the possibility of parallel acquisition of cyclic depsipeptide synthases in ancient complexes within Fusarium genus. Different fungi have independently evolved NRPS genes involved in depsipeptide biosynthesis, with functional adaptation towards biosynthesis of overlapping yet diversified metabolite profiles.

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Evolutionary Relationship between Chlorocatechol Catabolic Enzymes from Rhodococcus opacus 1CP and Their Counterparts in Proteobacteria: Sequence Divergence and Functional Convergence

Journal of Bacteriology ◽

10.1128/jb.180.5.1082-1094.1998 ◽

1998 ◽

Vol 180 (5) ◽

pp. 1082-1094 ◽

Cited By ~ 55

Author(s):

Dirk Eulberg ◽

Elena M. Kourbatova ◽

Ludmila A. Golovleva ◽

Michael Schlömann

Keyword(s):

Gene Cluster ◽

Regulatory Protein ◽

Evolutionary Relationship ◽

Sequence Divergence ◽

Functional Adaptation ◽

Open Reading Frames ◽

Rhodococcus Opacus ◽

Sequence Comparisons ◽

Catabolic Enzymes ◽

Chloromuconate Cycloisomerase

Biochemical investigations of the muconate and chloromuconate cycloisomerases from the chlorophenol-utilizing strainRhodococcus opacus (erythropolis) 1CP had previously indicated that the chlorocatechol catabolic pathway of this strain may have developed independently from the corresponding pathways of proteobacteria. To test this hypothesis, we cloned the chlorocatechol catabolic gene cluster of strain 1CP by using PCR with primers derived from sequences of N termini and peptides of purified chlorocatechol 1,2-dioxygenase and chloromuconate cycloisomerase. Sequencing of the clones revealed that they comprise different parts of the same gene cluster in which five open reading frames have been identified. The clcB gene for chloromuconate cycloisomerase is transcribed divergently from a gene which codes for a LysR-type regulatory protein, the presumed ClcR. Downstream of clcRbut separated from it by 222 bp, we detected the clcA andclcD genes, which could unambiguously be assigned to chlorocatechol 1,2-dioxygenase and dienelactone hydrolase. A gene coding for a maleylacetate reductase could not be detected. Instead, the product encoded by the fifth open reading frame turned out to be homologous to transposition-related proteins of IS1031 and Tn4811. Sequence comparisons of ClcA and ClcB to other 1,2-dioxygenases and cycloisomerases, respectively, clearly showed that the chlorocatechol catabolic enzymes of R. opacus 1CP represent different branches in the dendrograms than their proteobacterial counterparts. Thus, while the sequences diverged, the functional adaptation to efficient chlorocatechol metabolization occurred independently in proteobacteria and gram-positive bacteria, that is, by functionally convergent evolution.

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The possibilities of biologically protecting plants against diseases in nurseries, with special consideration of Oomycetes and Fusarium fungi

Forest Research Papers ◽

10.2478/frp-2014-0029 ◽

2014 ◽

Vol 75 (3) ◽

pp. 301-321 ◽

Cited By ~ 1

Author(s):

Adam Okorski ◽

Tomasz Oszako ◽

Justyna A. Nowakowska ◽

Agnieszka Pszczółkowska

Keyword(s):

Integrated Pest Management ◽

Economic Impact ◽

Pest Management ◽

Plant Protection ◽

Mechanisms Of Action ◽

Fusarium Spp ◽

High Quality ◽

Research Teams ◽

Forest Nurseries ◽

Fusarium Fungi

Abstract Achieving high quality propagative material is difficult today due to the limited number of pesticides recommended for use. Simultaneously, EU regulations on Integrated Pest Management (IPM) in forest nurseries came into a force, requiring a search for alternative plant protection methods that are safe for humans, animals and the environment. In this paper, we present the possibilities of using bio-fungicides against diseases in forest nurseries, their mechanisms of action, as well as the direction of their development (according to IPM rules). We reviewed the results achieved by different research teams presenting the possibilities and trends in combatting Oomycetes and Fusarium spp. pathogens currently having the most important economic impact.

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Direct measurement of electron-diffraction-pattern intensities using an energy loss spectrometer

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100155311 ◽

1989 ◽

Vol 47 ◽

pp. 668-669

Author(s):

A. G. Jackson ◽

M. Rowe

Keyword(s):

Thermal Effects ◽

Dynamic Range ◽

Scattering Amplitude ◽

Dynamical Theory ◽

Site Symmetry ◽

Proof Of Concept ◽

Parallel Acquisition ◽

Kinematic Approximation ◽

Speed Up ◽

Structure Calculations

Diffraction intensities from intermetallic compounds are, in the kinematic approximation, proportional to the scattering amplitude from the element doing the scattering. More detailed calculations have shown that site symmetry and occupation by various atom species also affects the intensity in a diffracted beam. [1] Hence, by measuring the intensities of beams, or their ratios, the occupancy can be estimated. Measurement of the intensity values also allows structure calculations to be made to determine the spatial distribution of the potentials doing the scattering. Thermal effects are also present as a background contribution. Inelastic effects such as loss or absorption/excitation complicate the intensity behavior, and dynamical theory is required to estimate the intensity value.The dynamic range of currents in diffracted beams can be 104or 105:1. Hence, detection of such information requires a means for collecting the intensity over a signal-to-noise range beyond that obtainable with a single film plate, which has a S/N of about 103:1. Although such a collection system is not available currently, a simple system consisting of instrumentation on an existing STEM can be used as a proof of concept which has a S/N of about 255:1, limited by the 8 bit pixel attributes used in the electronics. Use of 24 bit pixel attributes would easily allowthe desired noise range to be attained in the processing instrumentation. The S/N of the scintillator used by the photoelectron sensor is about 106 to 1, well beyond the S/N goal. The trade-off that must be made is the time for acquiring the signal, since the pattern can be obtained in seconds using film plates, compared to 10 to 20 minutes for a pattern to be acquired using the digital scan. Parallel acquisition would, of course, speed up this process immensely.

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